Acquired immune deficiency syndrome (AIDS) is an enormous health threat. (6). The causative agent on AIDS is the HIV-1 virus (6). Consequently, there is an ongoing effort towards identifying novel anti-HIV-1 therapeutics. One potent anti-HIV-1 target is the virally-encoded protease (Prt), which is essential for HIV-1 growth and replication (12, 20). The identification of Prt inhibitory compounds requires safe and reproducible Prt assays, which may be used to screen potential Prt-inhibitory compounds and to study the action of known Prt inhibitors. In this study, we propose the development of a superior assay, one that is safer, simpler, and less expensive than the currently available assays for Prt. The assay would measure Prt activity in vivo in a human T cell line by monitoring the activity of beta-galactosidase containing a Prt cleavage site. In addition to the ease and relevance of the proposed in vivo assay, its most valuable feature is that it involves no viruses or virus particles. To date, all Prt assays done in mammalian cells expose the worker to these agents. The long term goal is to develop this assay as a screen for anti-Prt compounds and to glean information useful for rational design of better therapeutics.